Oxygen gradients for open well cellular cultures via microfluidic substrates.
Oxygen gradients for open well cellular cultures via microfluidic substrates.
复制标题
通过微流体底物的开放良好细胞培养物的氧梯度。
DOI：
10.1039/c004660d
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发表时间：
2010-09-21
期刊：
影响因子：
6.1
通讯作者：
Eddington DT
中科院分区：
文献类型：
--
作者：
Lo JF;Sinkala E;Eddington DT
在微尺度上控制氧浓度有助于氧化应激、缺血和活性氧(ROS)介导的细胞通路的实验研究。在这里，我们报告了微流控梯度产生在开井培养模型中的应用，在该模型中，通过将细胞与下面的气体微通道分开的气体渗透基质的扩散来输送气体梯度。通过使用扩散来定位氧气输送，可以快速和可控地施加氧气浓度的微梯度，而不会使细胞暴露在机械压力下或减少微流控培养室内的培养体积。此外，我们还演示了应用这些氧气微梯度对Madin-Darby犬肾(MDCK)细胞内ROS水平的调节。在MDCK细胞中观察到与氧化应激和低氧暴露相一致的ROS水平的增加。在对照对照中，测量的ROS增加与100μM过氧化氢暴露相当，这在标准ROS诱导方法的范围内。与200μ的M孵育，维生素C能够解调在低氧和高氧暴露下的ROS反应。通过提供微流体控制的梯度、恒定的ROS暴露和无剪切开井设计，这里介绍的设备大大改进了标准的基于氧气的培养方法。这里介绍的设备可以在快速的时间尺度上产生复杂的氧气梯度，允许对许多难以建模的生理系统进行研究。
Controlling oxygen concentration at a microscale level can benefit experimental investigations involving oxidative stress, ischemia, and reactive oxygen species (ROS) mediated cellular pathways. Here, we report the application of microfluidic gradient generation in an open-well culture model, in which a gradient of gas is delivered via diffusion through a gas permeable substrate that separates cells from the gas microchannels below. By using diffusion to localize oxygen delivery, microgradients of oxygen concentrations can be rapidly and controllably applied without exposing cells to mechanical stresses or reducing culture volumes inside microfluidic culture chambers. Furthermore, we demonstrate the modulation of intracellular ROS levels in Madin-Darby canine kidney (MDCK) cells by applying these oxygen microgradients. Increases in ROS levels consistent with both oxidative stress and hypoxic exposures were observed in MDCK cells. The measured ROS increases were comparable to 100 μM hydrogen peroxide exposure in a control comparison, which is within the range of standard ROS induction methods. Incubation with 200 μM vitamin C was able to demodulate the ROS response at both hypoxic and hyperoxic exposures. By providing microfluidic controlled gradients, constant ROS exposure, and a shear-free open well design, the devices introduced here greatly improve upon standard oxygen-based culturing methods. The devices presented here can generate complex oxygen gradients over rapid timescales, permitting investigation of a number of difficult-to-model physiological systems.
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影响因子：
6.1
作者：
Adler M;Polinkovsky M;Gutierrez E;Groisman A
通讯作者：
Groisman A
影响因子：
6.1
作者：
Abate, Adam R.;Lee, Daeyeon;Weitz, David A.
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Weitz, David A.
影响因子：
--
作者：
Khan, SR
通讯作者：
Khan, SR
影响因子：
7.4
作者：
Khand, FD;Gordge, MP;Hothersall, JS
通讯作者：
Hothersall, JS
影响因子：
7.4
作者：
Kane, Bartholomew J.;Zinner, Michael J.;Toner, Mehmet
通讯作者：
Toner, Mehmet